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A kind of cd3ak cell culture composition and culture method thereof

A technology of cell culture and composition, which is applied in the field of cell culture, can solve the problem of low cell killing activity, and achieve the effects of reducing production costs, improving killing activity, and high purity

Active Publication Date: 2018-09-11
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are certain defects in this method. For example, although the number of cell expansion is large, the killing activity of the cells is low.

Method used

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  • A kind of cd3ak cell culture composition and culture method thereof
  • A kind of cd3ak cell culture composition and culture method thereof
  • A kind of cd3ak cell culture composition and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Culture of CD3AK cells

[0055] 1. Isolation of peripheral blood mononuclear cells

[0056] Collect 50-100mL of human peripheral blood, mix it with normal saline at a volume ratio of 1:1, and slowly add the mixed blood mixture and lymphocyte separation solution at a volume ratio of 2:1 to The upper layer of the lymphocyte separation solution was centrifuged at 3000rpm for 20min;

[0057] After centrifugation, from top to bottom of the centrifuge tube, they are plasma, buffy coat (i.e. mononuclear cell layer PBMC), lymphocyte separation medium, red blood cell layer, extract the PBMC layer, and wash and resuspend the collected PBMC with normal saline Afterwards, centrifuge at 1500rpm for 10min, repeat washing twice, and collect PBMC;

[0058] 2. Expansion and culture of CD3AK cells

[0059] Resuspend the collected PBMC with 1640 medium, press 1×10 6 Inoculate the culture flask at a density of 10 μg / mL, 100 U / mL and 10 U / mL; add 2% autologous Serum (autologo...

Embodiment 2

[0061] Example 2 Culture of CD3AK cells

[0062] 1. Isolation of peripheral blood mononuclear cells

[0063] Collect 50-100mL of human peripheral blood, mix it with normal saline at a volume ratio of 1:1, and slowly add the mixed blood mixture and lymphocyte separation solution at a volume ratio of 2:1 to The upper layer of the lymphocyte separation solution was centrifuged at 3000rpm for 20min;

[0064] After centrifugation, from top to bottom of the centrifuge tube, they are plasma, buffy coat (i.e. mononuclear cell layer PBMC), lymphocyte separation medium, red blood cell layer, extract the PBMC layer, and wash and resuspend the collected PBMC with normal saline Afterwards, centrifuge at 1500rpm for 10min, repeat washing twice, and collect PBMC;

[0065] 2. Expansion and culture of CD3AK cells

[0066] Resuspend the collected PBMC with 1640 medium, press 1×10 6 Inoculate the culture flask at a density of 50 μg / mL, 300 U / mL and 30 U / mL; add 20% autologous Serum (autolog...

Embodiment 3

[0068] Example 3 Culture of CD3AK cells

[0069] 1. Isolation of peripheral blood mononuclear cells

[0070] Collect 50-100mL of human peripheral blood, mix it with normal saline at a volume ratio of 1:1, and slowly add the mixed blood mixture and lymphocyte separation solution at a volume ratio of 2:1 to The upper layer of the lymphocyte separation solution was centrifuged at 3000rpm for 20min;

[0071] After centrifugation, from top to bottom of the centrifuge tube, they are plasma, buffy coat (i.e. mononuclear cell layer PBMC), lymphocyte separation medium, red blood cell layer, extract the PBMC layer, and wash and resuspend the collected PBMC with normal saline Afterwards, centrifuge at 1500rpm for 10min, repeat washing twice, and collect PBMC;

[0072] 2. Expansion and culture of CD3AK cells

[0073] Resuspend the collected PBMC with 1640 medium, press 1×10 6 Inoculate the culture flask at a density of 30 μg / mL, 200 U / mL and 20 U / mL; add 10% autologous Serum (autolog...

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Abstract

The invention relates to the technical field of cell culture, particularly to a CD3AK (anti-CD3antibody induced activated killer) cell culture composition and a culture method. The CD3AK cell culture composition comprises CDMcAb, IL-2, IL-15, pseudo-ginseng saponin, wogonin, blood serum and a basal culture medium. The CD3AK cell culture composition can promote mass proliferation of CD3AK cells, improves the killing activity of the CD3AK cells, avoids extrinsic protein and virus pollution, and greatly reduces production cost; the cultured CD3AK cells are relatively high in purity.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a CD3AK cell culture composition and a culture method thereof. Background technique [0002] CD3AK cells (anti-CD3antibody induced activated killer cells) refer to the killer cells of mononuclear cells such as lymphocytes activated by anti-CD3 monoclonal antibody (CD3McAb). -2 can maintain its active proliferation, so CD3 + CD8 + The T-dominated heterogeneous cell population has abundant sources of precursor cells, mainly T cells, which have a long survival time and low dependence on IL-2. [0003] CD3AK cells have broad-spectrum anti-tumor activity, and their pseudopods and protrusions are closely combined with target cells to kill tumor cells. The death forms of target cells are apoptosis and lytic necrosis. After TIL and TIL, the most concerned tumor effector cells in tumor AIT research. CD3AK cells have stronger expansion and anti-tumor ability than LAK cells and TIL...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
Inventor 陈海佳王一飞葛啸虎李丽娟
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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